Fig. 4

PHLPP1 and PHLPP2 as target genes of miR-224 in ESCC cells. a Predicted miR-224 target sequences in PHLPP1 and PHLPP2 3′-UTRs. The mutated nucleotides in PHLPP1 and PHLPP2 3′-UTRs used for our experiments are highlighted in red. b Luciferase assay. Interaction of miR-224 with 3′-UTR of PHLPP1 and PHLPP2 was confirmed by the luciferase assay in Eca109 and TE13 cells. c qRT-PCR. The data showed that miR-224 did not affect expression of PHLPP1 and PHLPP2 mRNA in Eca109 and TE13 cells (p > 0.05). Error bars represent the mean ± SD of three independent experiments. miR-224 M, miR-224 mimic; miR-224I, miR-224 inhibitor; NC, negative control; *p < 0.05 and **p < 0.01